2005 Annual Report 1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? What does it matter? Harvest of fish from the oceans has met or exceeded sustainable levels for all commercially important species, with the possible exception of the Alaskan fishery. Seafood consumption, however, is continuing to increase due to the influence of aquaculture production. It is projected that the output of commercial aquaculture will have to increase by 500% in the next 25 years to meet this growing demand. Fish meal and fish oil have been primary ingredients of fish feeds, but the finite supply of these crucial ingredients will begin to limit industry expansion within the next 10 years (Tacon, 2003). These "capture fisheries" products are also costly, variable in quality and availability, and considered by some to be unsustainable. To allow for the continued expansion of aquaculture, production efficiencies must be increased. Developing feeds that have fish meal and fish oil replaced with plant derived ingredients can reduce both fish feed cost and cost variability. The use of plant ingredients in fish feeds does create problems with palatability and digestibility of the feed, and growth rates are reduced. Open-formula vitamin premixes for trout have not been updated for the last 15 years despite considerable research on specific vitamin requirements. In addition, plant based feeds may require increased supplementation of specific vitamins involved with carbohydrate metabolism. Growth rate of fish may be impaired if adequate vitamin premixes are not developed. Also, current formulations do not take into account amino acid availabilities from the growing number of plant-derived ingredients, and this information is critical to optimize growth and nitrogen retention. The ratio of essential amino acids to one another that promotes optimal growth at the lowest protein levels is considered the ideal amino acid profile, and has been used successfully with other production animals. Combining an ideal amino acid profile and amino acid availability data for plant-derived ingredients will add flexibility and accuracy in feed formulation and help to maintain high growth performance of fish fed the improved feeds. Identifying factors responsible for growth rate reduction and developing solutions to these nutritional problems is required if plant based feeds are to be accepted by the fish farming industry. In nature trout have evolved as carnivores/insectivores, so distinct genetic stocks must be generated to optimize the growth efficiency of trout fed plant-based feeds. Thus, an additional and complementary approach to improving production efficiency is to develop strains of rainbow trout with an improved ability to use plant-based feeds. Recent research has indicated that rainbow trout strains selected for utilization of plant-based feeds perform better than unselected parental strains. For aquaculture to continue its growth, genetically enhanced domesticated stocks of fish that can utilize the plant-based feeds of the future will be needed. For aquaculture to continue its growth, genetically enhanced domesticated stocks of fish will need to be generated that are specific for the consumption of the most cost efficient and readily available diet. A successful conclusion to this research will enable the total replacement of fishmeal in trout feeds with sustainable plant-derived ingredients, while maintaining optimal production efficiencies. Attaining these objectives will provide economic, environmental, and health benefits to several sectors of society. Commercial fish producers will benefit through improved production efficiencies by using superior feeds and enhanced genetic strains. The use of grain-based feeds will enhance the sustainability of aquaculture by lessening dependence on marine protein and oils. Feed manufacturers will benefit by the availability of alternate feed ingredients, increased knowledge of specific nutrient requirements and access to proven feed formulations. Grain farmers will benefit by a new market demand for their products. Consumers will benefit by having affordable, high quality protein sources available that are produced in an environmentally sustainable manner because of reduced nutrient levels (phosphorus) in surface waters from aquaculture production. An additional benefit is that the reduction or elimination of marine based feedstuffs in trout feeds will remove pressure on currently over-exploited wild fishery stocks. This project is part of NP106 Aquaculture, and contributes to the action plan in 2 of the 5 areas identified. This project contributes to the development of brood stock/genetic improvement programs for carnivorous fish (trout). The project is also developing comprehensive multi-disciplinary research on the nutrient requirements of trout to allow them to perform efficiently when fed plant sources of nutrients. 2.List the milestones (indicators of progress) from your Project Plan. Milestone 1 (18 months) - a) Feed Development; Evaluation of processing on ingredients b) Nutrient Utilization; fatty acid requirement determinations Milestone 2 (24 months)- a) Feed Development; alternate oil evaluation Milestone 3 (48 months)- a) Feed Development; biological modification of ingredients b) Feed Development; mechanical fractionization of ingredients c) Nutrient Supplementation; vitamin requirements d) Nutrient Supplementation; amino acid requirements e) Nutrient Utilization; carbohydrate metabolism f) Nutrient Utilization; amino acid metabolism g) Selection Program; identification of markers Milestone 4 (60 months)- a) Feed Development; digestibility determinations b) Feed Development; complete feed development c) Selection Program; selective breeding program 4a.What was the single most significant accomplishment this past year? Understanding the genetic components in trout involved in diet utilization is critical for US aquaculture industry to remain competitive particularly in the face of higher feed costs, due limited natural and protein resources, and stricter environmental standards. Scientists in the Small Grains and Potato Germplasm Research Unit, Aberdeen, ID. studied differential expression relating to trout growth, health and metabolism reared on formulated cereal grain and commercial feeds. Rainbow trout of distinct lineage were separated and reared on either a commercial diet or a diet containing a high level of barley (32%) for one year after which, tissue samples were taken and the changes in gene expression were evaluated through analysis using quantitative real-time PCR for several genes and microarray hybridization. The findings from this work will now allow researchers more efficient means to evaluate selected broodstock for improved performance and aid in the evaluation of formulated diets. 4b.List other significant accomplishments, if any. Mixtures of plant-products were fermented using Solid Substrate Culture to reduce anti-nutrients and increase the concentration of nutrients. Using laboratory culture apparatus, mixtures of full fat soybeans and barley were fermented with specific micro-organism and the level of crude protein was increased from approximately 35% to 58%. Each ingredient was added to a semi-purified control diet containing an inert, marker compound (yttrium oxide) and fed to post-juvenile rainbow trout. Feces were collected and analyzed, along with each ingredient, test diets containing each ingredient, and the control diet, and apparent digestibility coefficients (ADC) for dry matter, organic matter, and protein, lipid, energy and phosphorus were calculated. The ADC for organic matter, protein and lipid digestibility's increased from 51.4, 57.3 and 74.0 to 88.6, 97.4 and 109.6%, respectively as a result of the fermentation, and ADC for energy and phosphorus also increased from 45.7 and 84.6 to 91.1 and 100% respectively. The improved nutrient digestibility of these products indicate potential for replacing fish meal, nutrients in the effluent waters and reducing fecal output of trout. Taurine was identified as a previously unidentified growth factor present in fishmeal that is not present in plant derived ingredients. Research has demonstrated that taurine supplementation is required when rainbow trout are fed diets containing protein form only plant derived ingredients. The supplementation of taurine to these diets improves growth rates, feed conversion efficiencies, protein retention efficiencies and energy retention efficiencies. 4c.List any significant activities that support special target populations. Lectures in aquaculture class for Native Americans by Overturf, Barrows and Gaylord. 4d.Progress report. None. 5.Describe the major accomplishments over the life of the project, including their predicted or actual impact. This is a newly initiated project and was started as a joint collaboration between the Small Grains and Potato Germplasm Facility in Aberdeen, Idaho and the Hagerman Fish Culture Experiment Station in Hagerman, Idaho. Please refer to the report of previous project 5366-21310-002-00D. With this being a new and unique area of research within the Agricultural Research Service, there has been rapid progress in development of plant-derived ingredients, nutrient requirement and availability information and in improving trout strains for growth and the utilization of these new diets. During the span of this project we plan to develop three categories of products. First, improved ingredients will be identified and developed for use in trout feeds. Second, feed formulations and nutrient supplement specifications for plant-based feeds will be developed. Third, genes involved with nutrient metabolism and utilization of trout when fed plant-based feeds will be identified and characterized, and improved genetic strains of rainbow trout will be selected utilizing this information. Trout farmers, fish feed manufacturers, grain producers and consumers will all use products from this project. Scientists studying genetic improvement of rainbow trout will employ genetic markers for traits associated with metabolism of plant products by trout. Consumers demanding safe aquaculture products will enjoy healthy products produced in an environmentally responsible manner. 6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products? This project is replacing 5366-21310-002-00D per OSQR review. 7.List your most important publications in the popular press and presentations to organizations and articles written about your work. (NOTE: List your peer reviewed publications below). Barrows, F., Bradley, C., Sands, D., and Kirkpatrick, M. (2005) Methods for improving the nutrient profile of plant-derived ingredients. World Aquaculture Society, New Orleans, LA Barrows, F., Overturf, K., Gaylord, G., and Bonman M. (2005) Improving production efficiency of rainbow trout (Oncorhynchus mykiss). National Center for Cool and Coldwater Aquaculture, Leetown, WV. Barrows F. (2005) Plant-derived ingredients for aquaculture feeds; biological concentration and mechanical fractionization. Special Workshop hosted by United Soybean Board, Twin Falls, Idaho Barrows F. (2005). Feed ingredients; old problems and new approaches. U.S. Trout Farmers Meeting, New Orleans, LA Barrows, F., Overturf, K., Gaylord, G., and Bonman M. (2005) The integration of nutritional, genetic and physiological approaches to improve production efficiency of rainbow trout (Oncorhynchus mykiss). Crop Conversion Science and Engineering Research Unit, ERRC, Wyndmor, PA Johansen, K. and Overturf K. (2005) Quantitative expression analysis of genes affecting muscle growth during the development of rainbow trout Onchorhynchus mykiss. World Aquaculture Society, New Orleans, LA Stone, D., Overturf, K. and Hardy, R. (2005) Can spray dried porcine plasma or whey concentrate reduce soybean meal induced enteritis in rainbow trout Onchorhynchus mykiss. World Aquaculture Society, New Orleans, LA Overturf, K., Rise, M. and Casten, M. (2005) Microarray expression differences in rainbow trout reared on an alternative or commercial diets. World Aquaculture Society, New Orleans, LA Overturf, K. (2005) Experimental evaluation program for USDA Rainbow Trout in Hagerman, Idaho. Idaho Aquaculture Association, Twin Falls, ID Overturf, K. (2005) Relating genetics and plant utilization in aquaculture. US Soybean Meeting, Twin Falls, Idaho Overturf, K. (2005) Correlating gene expression with physiological changes in rainbow trout. Idaho Bioinformatics Conference, Pocatello, Idaho Gaylord, T.G., A. M. Teague and F. T. Barrows. 2005 Taurine supplementation for rainbow trout (Oncorhynchus mykiss). Idaho Aquaculture Association Annual Meeting. Twin Falls, ID. Gaylord, T.G., A. M. Teague and F. T. Barrows. 2005 Taurine supplementation for rainbow trout (Oncorhynchus mykiss). Workshop on Plant Based Products in Aquafeeds. Twin Falls, ID. Davis, K.B. and T.G. Gaylord. 2005. Metabolic consequences of feed deprivation in sunshine bass. Aquaculture America 2005. New Orleans, LA. Rawles, S.D., T.G. Gaylord, and R. Lochmann. 2005. Glucose oxidation and lipogenesis in hybrid striped bass fed diets with different starch ratios. Joint meeting of the federation of Animal Science Societies. Cincinnati, OH. Rawles, S.D., T.G. Gaylord, and R. Lochmann. 2005. Hepatic glucose oxidation and lipogenesis in sunshine bass fed diets with different amylose to amylopectin ratios. Aquaculture America 2005. New Orleans, LA. Lochmann, R., S.D. Rawles, and T.G. Gaylord. 2005. Body indices, blood lipid class composition, and proximate analysis of sunshine bass fed diets with different amylose to amylopectin ratios. Aquaculture America 2005. New Orleans, LA. Review Publications Overturf, K.E., Cain, K.D., Cavender, W.P., Powell, M.S., Wood, J.S. 2004. Real-time quantitative polymerase chain reaction (qpcr) to identify myxobolus cerebralis in rainbow trout oncorhynchus mykiss. Diseases of Aquatic Organisms. v.60.p.205-213. Biga, P.R., Cain, K.D., Hardy, R.W., Shelling, G.T., Overturf, K., Roberts, S.B., Goetz, F.W. and Ott, T.L. Growth hormone differentially regulates muscle myostatin1 and -2 and increases circulating cortisol in rainbow trout (Oncorhynchus mykiss). (2004) General and Comparative Endocrinology 138: 32-41. Lellis, W.A., F.T. Barrows, and R.W. Hardy. 2004. Effects of phase-feeding dietary phosphorus on survival, growth, and processing characteristics of rainbow trout Oncorhynchus mykiss (Walbaum). Aquaculture. 224; 607-616. Liu, K.K.M., F.T. Barrows, R.W. Hardy, F.M. Dong. 2004. Body Composition, growth performance and product quality of rainbow trout (Onchorynchus mykiss) fed diets containing poultry fat, soybean/corn lecithin, or menhaden oil. Aquaculture. 238:309-328 Gaylord, T.G., Rawles, S.D., Gatlin, D.M. 2004. Amino acid availability from animal, blended, and plant feedstuffs for hybrid striped bass (Morone chrysops x M. saxatilis). Aquaculture Nutrition. 10(5):345-352.